Compound-specific Stable Carbon Isotope Analysis Research Articles

Two-dimensional high performance liquid chromatography purification of underivatized urinary prednisone and prednisolone for compound-specific stable carbon isotope analysis.

The gas chromatography-combustion isotope ratio mass spectrometry (GC/C/IRMS) confirmation procedure for prednisone (PS) and prednisolone (PSL) is still a great challenge for the doping control laboratory due to the many structurally similar steroids present in urinary matrices. This study aims to establish an innovative online two-dimensional high performance liquid chromatography (2D-HPLC) purification method for measuring the carbon isotope ratios (CIRs) and achieving the identification of the synthetic forms of these two endogenous anabolic androgenic steroids (EAASs). Initially, the one-dimensional chromatographic column was used to separate and purify endogenous reference compounds (ERCs), and the co-elution fluids containing PS and PSL were switched to a two-dimensional chromatographic column for further purification through an online transfer system. Then the purified compounds were analyzed using GC/C/IRMS after sample pretreatments. The results showed that the minimum detection concentration of PS and PSL reached 30 ng mL-1, and no isotope fractionation occurred during the entire collection and preparation process. This method has been validated with the WADA technical document and showed good sensitivity and selectivity, demonstrating its practical applicability for urine samples in doping control laboratories.

Transformation of Iminodi(methylene phosphonate) on Manganese Dioxides - Passivation of the Mineral Surface by (Formed) Mn2.

Aminopolyphosphonates (APPs) are strong chelating agents with growing use in industrial and household applications. In this study, we investigated the oxidation of the bisphosphonate iminodi(methylene phosphonate) (IDMP) - a major transformation product (TP) of numerous commercially used APPs and a potential precursor for aminomethylphosphonate (AMPA) - on manganese dioxide (MnO2). Transformation batch experiments at pH 6 revealed AMPA and phosphate as main TPs, with a phosphorus mass balance of 80 to 92% throughout all experiments. Our results suggest initial cleavage of the C-P bond and formation of the stable intermediate N-formyl-AMPA. Next, C-N bond cleavage leads to the formation of AMPA, which exhibits lower reactivity than IDMP. Reaction rates together with IDMP and Mn2+ sorption data indicate formation of IDMP-Mn2+ surface bridging complexes with progressing MnO2 reduction, leading to the passivation of the mineral surface regarding IDMP oxidation. Compound-specific stable carbon isotope analysis of IDMP in both sorbed and aqueous fractions further supported this hypothesis. Depending on the extent of Mn2+ surface concentration, the isotope data indicated either sorption of IDMP to the mineral surface or electron transfer from IDMP to MnIV to be the rate-limiting step of the overall reaction. Our study sheds further light on the complex surface processes during MnO2 redox reactions and reveals abiotic oxidative transformation of APPs by MnO2 as a potential process contributing to widespread elevated AMPA concentrations in the environment.

An Overview of Lipid Biomarkers in Terrestrial Extreme Environments with Relevance for Mars Exploration.

Lipid molecules are organic compounds, insoluble in water, and based on carbon-carbon chains that form an integral part of biological cell membranes. As such, lipids are ubiquitous in life on Earth, which is why they are considered useful biomarkers for life detection in terrestrial environments. These molecules display effective membrane-forming properties even under geochemically hostile conditions that challenge most of microbial life, which grants lipids a universal biomarker character suitable for life detection beyond Earth, where a putative biological membrane would also be required. What discriminates lipids from nucleic acids or proteins is their capacity to retain diagnostic information about their biological source in their recalcitrant hydrocarbon skeletons for thousands of millions of years, which is indispensable in the field of astrobiology given the time span that the geological ages of planetary bodies encompass. This work gathers studies that have employed lipid biomarker approaches for paleoenvironmental surveys and life detection purposes in terrestrial environments with extreme conditions: hydrothermal, hyperarid, hypersaline, and highly acidic, among others; all of which are analogous to current or past conditions on Mars. Although some of the compounds discussed in this review may be abiotically synthesized, we focus on those with a biological origin, namely lipid biomarkers. Therefore, along with appropriate complementary techniques such as bulk and compound-specific stable carbon isotope analysis, this work recapitulates and reevaluates the potential of lipid biomarkers as an additional, powerful tool to interrogate whether there is life on Mars, or if there ever was.

Mechanistic investigation of phosphonate photolysis in aqueous solution by simultaneous LC-IRMS and HRMS analysis

The mechanism of photolytic degradation of aminotrimethylphosphonic acid (ATMP) in an aqueous solution is not completely elucidated with respect to the transformation products formed during technical photodegradation. For the first time, the degradation of ATMP was investigated with simultaneous analysis by isotope ratio and high-resolution mass spectrometry (LC-IRMS+HRMS). This instrumental coupling combines quantitative data on degradation with compound-specific stable carbon isotope analysis (CSIA) in terms of kinetic isotope effects while identifying transformation products. Additionally, in combination with wet chemical analysis of ortho-phosphate (o-PO4), phosphorus balances were established to elucidate and interpret the degradation mechanism of ATMP under photolysis. Various experimental conditions during photolysis (temperature, dissolved oxygen concentration, and wavelength range) were investigated.ATMP was transformed to iminodimethylphosphonic acid (IDMP), aminomethylphosphonic acid (AMPA), and o-PO4 as main products during photolysis with a medium pressure mercury lamp. UV radiation < 310 nm was found to be required for ATMP degradation. The degradation of ATMP and its transformation products is slightly affected by temperature and dissolved oxygen concentration. Considering possible reactive oxygen species, such as hydroxyl radicals, it is assumed that the degradation of ATMP occurs to a small extent via a parallel degradation pathway in addition to photolysis. Complementary CSIA revealed an inverse carbon isotopic effect of ATMP due to direct photolysis, unaffected by the experimental conditions considered, while no isotopic effect was associated with the hydroxyl radical (H2O2) oxidation of ATMP. In UV/H2O2 oxidation of ATMP, four of five additional unknown transformation products, which have not yet been described in literature, could be assigned to postulated structures based on HRMS data. The findings achieved in this study demonstrate the advantages of simultaneous analysis by LC-IRMS+HRMS as a tool to elucidate reaction mechanisms for the first time.

Late Paleocene CO&amp;lt;sub&amp;gt;2&amp;lt;/sub&amp;gt; drawdown, climatic cooling and terrestrial denudation in the southwest Pacific

Abstract. ​​​​​​​Late Paleocene deposition of an organic-rich sedimentary facies on the continental shelf and slope of New Zealand and eastern Australia has been linked to short-lived climatic cooling and terrestrial denudation following sea level fall. Recent studies confirm that the organic matter in this facies, termed “Waipawa organofacies”, is primarily of terrestrial origin, with a minor marine component. It is also unusually enriched in 13C. In this study we address the cause of this enrichment. For Waipawa organofacies and its bounding facies in the Taylor White section, Hawke's Bay, paired palynofacies and carbon isotope analysis of heavy liquid-separated density fractions indicate that the heaviest δ13C values are associated with degraded phytoclasts (woody plant matter) and that the 13C enrichment may be partly due to lignin degradation. Compound-specific stable carbon isotope analyses of samples from the Taylor White and mid-Waipara (Canterbury) sections display similar trends and further reveal a residual 13C enrichment of ∼ 2.5 ‰ in higher plant biomarkers (long chain n-alkanes and fatty acids) and a ∼ 2 ‰–5 ‰ change in subordinate marine biomarkers. Using the relationship between atmospheric CO2 and C3 plant tissue δ13C values, we determine that the 3 ‰ increase in terrestrial δ13C may represent a ∼ 35 % decrease in atmospheric CO2. Refined age control for Waipawa organofacies indicates that deposition occurred between 59.2 and 58.5 Ma, which coincides with an interval of carbonate dissolution in the deep sea that is associated with a Paleocene oxygen isotope maximum (POIM, 59.7–58.1 Ma) and the onset of the Paleocene carbon isotope maximum (PCIM, 59.3–57.4 Ma). This association suggests that Waipawa deposition occurred during a time of cool climatic conditions and increased carbon burial. This relationship is further supported by published TEX86-based sea surface temperatures that indicate a pronounced regional cooling during deposition. We suggest that reduced greenhouse gas emissions from volcanism and accelerated carbon burial, due to tectonic factors, resulted in short-lived global cooling, growth of ephemeral ice sheets and a global fall in sea level. Accompanying erosion and carbonate dissolution in deep-sea sediment archives may have hidden the evidence of this “hypothermal” event until now.

Two Pathways Compete in the Mn(II)-Catalyzed Oxidation of Aminotrismethylene Phosphonate (ATMP).

Mn(II)-catalyzed oxidation by molecular oxygen is considered a relevant process for the environmental fate of aminopolyphosphonate chelating agents such as aminotrismethylene phosphonate (ATMP). However, the potential roles of Mn(III)ATMP-species in the underlying transformation mechanisms are not fully understood. We combined kinetic studies, compound-specific stable carbon isotope analysis, and equilibrium speciation modeling to shed light on the significance of such Mn-ATMP species for the overall ATMP oxidation by molecular oxygen. The fraction of ATMP complexed with Mn(II) inversely correlated with both (i) the Mn(II)-normalized transformation rate constants of ATMP and (ii) the observed carbon isotope enrichment factors (εc-values). These findings provide evidence for two parallel ATMP transformation pathways exhibiting distinctly different reaction kinetics and carbon isotope fractionation: (i) oxidation of ATMP present in Mn(III)ATMP complexes (εc ≈ -10 ‰) and (ii) oxidation of free ATMP by such Mn(III)ATMP species (εc ≈ -1 ‰) in a catalytic cycle. The higher reaction rate of the latter pathway implies that aminopolyphosphonates can be trapped in catalytic Mn-complexes before being transformed and suggests that Mn(III)ATMP might be a potent oxidant also for other reducible solutes in aqueous environments.

Compound-specific stable carbon and hydrogen isotope analyses of late-Holocene vegetation and precipitation change at Laguna Los Mangos, Costa Rica

New compound-specific isotope analyses of a sediment core from Laguna Los Mangos in southern Pacific Costa Rica improves understanding of late-Holocene precipitation change in a region with limited paleoprecipitation records that is vulnerable to future climate change. We established paleoprecipitation and paleovegetation records from compound-specific stable hydrogen and carbon isotopic compositions of terrestrially-derived n-alkanes (δ2Halkane and δ13Calkane) to assess paleohydrologic variability and potential linkages to paleoecological change and human activity as revealed by prior analyses of the Los Mangos core. The δ2Halkane values were corrected for isotopic fractionation using pollen counts from the same core. The Los Mangos record extends to 4200 cal yr BP and small increases in δ13C values of C29, +0.6‰, and C31 alkanes, +0.3‰, (δ13CC29, 31) indicate a slight increase in C4 vegetation after initial introduction of maize agriculture to the watershed at ca. 3360 cal yr BP. This slight increase in C4 vegetation is followed by the largest positive carbon isotope excursions in the record, as compared to record averages (δ13CC29 = +3.2‰, δ13CC31 = +5.0‰). Paleohydrologic variability likely influenced vegetation and human activity at Los Mangos. Lake desiccation during the late-Terminal Classic Drought (TCD) resulted in a sedimentary hiatus in the Los Mangos record from ca. 950 to 450 cal yr BP. Positive excursions in comparison to record averages occur for both δ2HC29 and δ2HC31 proxies (δ2HC29 = +25.3 to +13.4‰ and δ2HC31 = +6.5‰) during the middle Little Ice Age (LIA) and indicate drier than average conditions, but there is no evidence of desiccation during this period. Thus, drought conditions during the LIA were apparently not as severe at Los Mangos as during the TCD, possibly because of differing forcing mechanisms for LIA climate that originated, or were more clearly expressed, in the Atlantic basin.

Evaluation of a quick one-step sample preparation method for the determination of the isotopic fingerprint of rapeseed (Brassica napus): Investigation of the influence of the use of 2,2-dimethoxypropane on compound-specific stable carbon and hydrogen isotope analyses by gas chromatography combustion/pyrolysis isotope ratio mass spectrometry.

Gas chromatographic analyses for vegetable oils require transesterification, which generally involves multiple steps, mainly to generate fatty acid methyl esters (FAMEs). A quick method based on acid-catalyzed transesterification using 2,2-dimethoxypropane (DMP) enables the conversion in one step, in a single reactor. For compound-specific stable carbon and hydrogen isotope analyses (C- and H-CSIA) of individual fatty acids (FAs) in oil, the verification of this one-step method has not yet been reported. In this study, we evaluated the feasibility of the one-step method for C- and H-CSIA of individual FAMEs in rapeseed samples. The focus was on the investigation of the influence of methanol, which was produced from the reactions of DMP with glycerol and water during transesterification, on the accuracy of isotope composition of FAMEs, consequently of the FAs. The reproducibility of the one-step method was assessed by the measurement of the FAMEs from rapeseed and rapeseed oil. For the C- and H-CSIA of individual FAMEs, a gas chromatography combustion/pyrolysis isotope ratio mass spectrometry system was used. Our results showed that no significant differences arise in the carbon and hydrogen isotope compositions of the selected main FAMEs produced with and without DMP except for the H-CSIA value of C18:3. The reproducibility of the one-step method for rapeseed was in the range of ±0.1 mUr to ± 0.3 mUr for C-CSIA and ±1 mUr to ±3 mUr for H-CSIA of the main FAMEs. DMP improves the transesterification efficiency without influencing the accuracy of the C- and H-CSIA of FAMEs. The performance of the one-step method for rapeseed samples for the determination of C- and H-CSIA values of FAMEs is satisfactory. Thus, the applicability of the one-step method for isotopic fingerprint analyses of FAs in oilseeds is reported for the first time.

In Situ Microbial Degradation of PBDEs in Sediments from an E-Waste Site as Revealed by Positive Matrix Factorization and Compound-Specific Stable Carbon Isotope Analysis.

In the present study, positive matrix factorization (PMF) and compound-specific isotope analysis were used to investigate the in situ biodegradation of polybrominated diphenyl ethers (PBDEs) in sediment cores collected from a pond at an e-waste recycling site in South China. The potential microorganisms relevant to the degradation of PBDEs were also assessed to aid in the understanding of in situ biodegradation. The PMF results suggested that reductive debromination took place in the sediments. The debromination signal (ratio of the concentration of factor 5 (PMF result) to the total PBDE content) was positively correlated with the relative abundance of Dehalococcoidetes at different core depths. The clear 13C enrichment of five PBDE congeners (BDE 28, 47, 49, 99, and 153) with increasing core depth indicated that a measurable change in isotope fractionation might have occurred during PBDE biodegradation. The in situ biodegradation was further validated by the widespread detection of mono-BDE congeners (BDE 2, BDE 3) and diphenyl ether in the sediments. This study provides new evidence to enhance our understanding of the in situ biodegradation of PBDEs and suggests that the extensive removal of bromine from PBDEs was mediated by indigenous microorganisms at the e-waste site.

Toluene biodegradation in the vadose zone of a poplar phytoremediation system identified using metagenomics and toluene-specific stable carbon isotope analysis

Biodegradation is an important mechanism of action of phytoremediation systems, but performance evaluation is challenging. We applied metagenomic molecular approaches and compound-specific stable carbon isotope analysis to assess biodegradation of toluene in the vadose zone at an urban pilot field system where hybrid poplars were planted to remediate legacy impacts to an underlying shallow fractured bedrock aquifer. Carbon isotope ratios were compared spatio-temporally between toluene dissolved in groundwater and in the vapor phase. Enrichment of 13C from toluene in the vapor phase compared to groundwater provided evidence for biodegradation in the vadose zone. Total bacterial abundance (16S rRNA) and abundance and expression of degradation genes were determined in rhizosphere soil (DNA and RNA) and roots (DNA) using quantitative PCR. Relative abundances of degraders in the rhizosphere were on average higher at greater depths, except for enrichment of PHE-encoding communities that more strongly followed patterns of toluene concentrations detected. Quantification of RMO and PHE gene transcripts supported observations of active aerobic toluene degradation. Finally, spatially-variable numbers of toluene degraders were detected in poplar roots. We present multiple lines of evidence for biodegradation in the vadose zone at this site, contributing to our understanding of mechanisms of action of the phytoremediation system.

Multi-method assessment of the intrinsic biodegradation potential of an aquifer contaminated with chlorinated ethenes at an industrial area in Barcelona (Spain)

The bioremediation potential of an aquifer contaminated with tetrachloroethene (PCE) was assessed by combining hydrogeochemical data of the site, microcosm studies, metabolites concentrations, compound specific-stable carbon isotope analysis and the identification of selected reductive dechlorination biomarker genes. The characterization of the site through 10 monitoring wells evidenced that leaked PCE was transformed to TCE and cis-DCE via hydrogenolysis. Carbon isotopic mass balance of chlorinated ethenes pointed to two distinct sources of contamination and discarded relevant alternate degradation pathways in the aquifer. Application of specific-genus primers targeting Dehalococcoides mccartyi species and the vinyl chloride-to-ethene reductive dehalogenase vcrA indicated the presence of autochthonous bacteria capable of the complete dechlorination of PCE. The observed cis-DCE stall was consistent with the aquifer geochemistry (positive redox potentials; presence of dissolved oxygen, nitrate, and sulphate; absence of ferrous iron), which was thermodynamically favourable to dechlorinate highly chlorinated ethenes but required lower redox potentials to evolve beyond cis-DCE to the innocuous end product ethene. Accordingly, the addition of lactate or a mixture of ethanol plus methanol as electron donor sources in parallel field-derived anoxic microcosms accelerated dechlorination of PCE and passed cis-DCE up to ethene, unlike the controls (without amendments, representative of field natural attenuation). Lactate fermentation produced acetate at near-stoichiometric amounts. The array of techniques used in this study provided complementary lines of evidence to suggest that enhanced anaerobic bioremediation using lactate as electron donor source is a feasible strategy to successfully decontaminate this site.

Compound-specific stable carbon isotope analysis of lignin phenols by gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS)

Compound-specific stable carbon isotope analysis of lignin phenols by gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS)

多環芳香族炭化水素類(PAHs)の安定炭素同位体分析を用いた環境化学的研究の現状と展望

多環芳香族炭化水素類(PAHs)の安定炭素同位体分析を用いた環境化学的研究の現状と展望

Labelled microbial culture as a calibration medium for (13) C-isotope measurement of derivatized compounds: application to tert-butyldimethylsilyl amino acids.

Compound-specific stable carbon isotope analysis by gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) is widely used in studies of environmental or biological functioning. In the case of derivatized molecules, a calibration might be required due to added non-analyte carbon and in some cases non-stoichiometric recovery by the mass spectrometer. Two biological materials of known isotopic composition were produced by microbial cell cultures on either (13) C-labelled glucose or non-labelled glucose as sole source of carbon. Subsequent hydrolyzed amino acids were derivatized as tert-butyldimethylsilyl (tBDMSi) derivatives and analyzed by GC/C/IRMS. The (13) C-enrichment measurements were used as a direct calibration to calculate the original (13) C/(12) C ratios of individual amino acids. We tested this calibration on both known and unknown samples. For the main proteinogenic amino acids we could determine the number of non-analyte added carbon atoms and assess the non-stoichiometrical recovery of tBDMSi carbon atoms, due to their incomplete oxidation in the combustion step of GC/C/IRMS. The calibration enabled the determination of the natural abundances (δ(13) C values) of amino acids with an average accuracy of ±1.1 ‰. We illustrate the application of the calibration to determine the (13) C/(12) C ratios of amino acids, and the associated uncertainty, in biological and plant materials. The analysis of a labelled microbial cell culture offers a straightforward, rapid and reliable estimate of non-analyte carbon contribution to stable isotope composition. We recommend this method as a calibration or a control in artificial or natural (13) C-tracing experiments. Copyright © 2016 John Wiley & Sons, Ltd.

Tracing the biotransformation of polychlorinated biphenyls (PCBs) in common carp (Cryprinus carpio): Enantiomeric fraction and compound-specific stable carbon isotope analyses

Metabolites of polychlorinated biphenyls (PCBs) in fish are difficult to detect in vivo due to the complexity of biometabolism. In the present study, atropisomeric fraction analysis of chiral PCB congeners and compound-specific isotopic analysis (CSIA) were applied to trace the biotransformation of PCBs in fish by exposure of common carp (Cryprinus carpio) to the commercial PCB mixture Aroclor 1242. Stereoselective elimination of the chiral PCB congeners 91, 95, and 136 was observed, indicating a stereoselective biotransformation process. The δ13C values of PCBs 5/8, 18, and 20/33 in fish were increased compared with those in the spiked food, while PCBs 47/48 and 49 showed significant heavy isotope depletion. These results suggested a significant biotransformation of the corresponding individual PCB congeners although the potential PCB metabolites, hydroxylated PCBs (OH-PCBs) and methylsulfone PCBs (MeSO2-PCBs), were not detected in the fish tissue samples throughout this experiment. The results of the present study demonstrated that a combination of chiral analysis and CSIA is a promising new approach for investigating the biotransformation of PCBs in biota.

Petroleum source, maturity, alteration and mixing in the southwestern Barents Sea: New insights from geochemical and isotope data

The Hammerfest Basin (HB) is an important area for petroleum exploration in the southwestern Norwegian Barents Sea. This contribution is aimed to characterize oils and condensates hosted in Triassic and Jurassic reservoirs, establish genetic relationships and to assess maturity, mixing and petroleum alteration processes. The research was based on geochemical as well as bulk and compound-specific stable carbon and hydrogen isotope analyses of oil and condensate samples from all major fields and discoveries in the HB, including extracts from Jurassic and Triassic source rocks. Interpretations of light hydrocarbon composition and isotope data clearly identify four main petroleum groups affected by microbial degradation, water washing, evaporative fractionation and high thermal maturity level. Based on polycyclic aromatic hydrocarbons (PAHs), marine and higher-plant-derived biomarkers and carbon isotopic composition (δ13C) of individual petroleum hydrocarbons, four petroleum families (I–IV) are recognized in the HB and a distinction can be made among oils and condensates from different stratigraphic levels and sectors of the Basin. Most oils and condensates from the HB consist of mixtures with small maturity variations; however, oils from Middle Triassic reservoirs in the Goliat field showed larger maturity variations according to biomarker, PAH and light hydrocarbon–maturity ratios. In the HB, geochemical and isotopic data allowed unraveling mixtures of hydrocarbons derived mainly from Jurassic, Triassic and Paleozoic source rocks.

Optimisation of solid-state urea clathrate formation as a chemical separation method coupled to compound-specific stable carbon isotope analysis

ABSTRACTSolid-state urea clathrate formation (SSUCF) as a chemical separation method prior to stable carbon isotope fingerprinting of diesel fuel contaminations was studied. The stable carbon isotope ratios (δ13C) of n-alkanes in diesel fuel can be used to trace the origin of a contamination. The accurate measurement of the stable isotopic composition of individual compounds requires baseline separation from any other co-eluting compounds. For this purpose silica gel column chromatography (SGCC) and SSUCF were applied. Detailed optimisation of SSUCF was performed: different activators, clathrate formation temperatures, activator volumes, clathrate formation times and sample capacity were investigated. The main benefits of the developed method are reduced clathrate formation time and increased recoveries for lower molecular weight n-alkanes. The recoveries of the developed SSUCF method ranged between 63 and 100% for C10–C24 n-alkanes with relative standard deviation no more than 7%. The precision of the gas chromatography-isotope ratio mass spectrometry measurement was acceptable with a standard deviation of the δ13C values ranging between 0.08 and 0.15‰. The absence of isotopic fractionation was also investigated.The robustness of the method was tested within a model experiment. Nine different water samples including distilled water, tap water, river water, industrial wastewaters and groundwater samples were spiked with the same diesel fuel. The water samples were extracted with n-hexane and after purification with both SGCC and SSUCF n-alkanes were measured. The δ13C values of n-alkanes were found to be similar for all samples. The importance of sample purification prior to compound-specific isotope analysis (CSIA) was also demonstrated within this model experiment by analysing samples from different stages of the sample preparation.Our results show that the proposed method can remarkably improve the precision of compound-specific stable carbon isotope analysis of n-alkanes originating from diesel contamination of the aquatic environment.

Compound-Specific Stable Carbon Isotope Analysis of Chlorofluorocarbons in Groundwater.

Chlorofluorocarbons (CFCs) and hydrochlorofluorocarbons (HCFCs), controlled substances due to their role in stratospheric ozone loss, also occur as dissolved contaminants in groundwaters. Stable carbon isotopic signatures may provide valuable new information on the fate of these compounds as has been seen for other priority hydrocarbon contaminants, but to date no method for extraction and isotopic analysis of dissolved CFCs from groundwaters has been developed. Here we describe a cryogenic purge and trap system coupled to continuous flow compound-specific stable carbon isotope analysis mass spectrometry for concentrations as low as 35 μg/L. The method is validated by comparing isotopic signatures from water extracted CFCs against a new suite of isotopic CFC standards. Fractionation of CFCs in volatilization experiments from pure-phase CFC-11 and CFC-113 resulted in enrichment factors (ε) of +1.7 ± 0.1‰ and +1.1 ± 0.1‰, respectively, indicating that such volatile loss, if significant, would produce a more (13)C depleted signature in the remaining CFCs. Importantly, no significant fractionation was observed during volatile extraction of dissolved CFCs from aqueous solutions. δ(13)C values for groundwaters from a CFC-contaminated site were, on average, more enriched than δ(13)C values for pure compounds. Such enriched δ(13)C values have been seen in other hydrocarbon contaminants such as chlorinated ethenes and ethanes due to in situ degradation, but definitive interpretation of such enriched signatures in field samples requires additional experiments to characterize fractionation of CFCs during biodegradation. The establishment of a robust and sensitive method of extraction and analysis, as described here, provides the foundation for such future directions.

Predominance of archaea-derived hydrocarbons in an Early Triassic microbialite

We investigated the distribution of lipids in Lower Triassic sedimentary rocks (252–247myr) from South China, including a shallow water microbialite in the uppermost section of the outcrop. Archaeal derived hydrocarbons were the major constituents of the microbialite from the latest Early Triassic. Among these, we detected (i) abundant C40 acyclic and monocyclic biphytanes (possibly derived from glycerol dialkyl glycerol tetraether lipids) and their degradation products, C30–39 pseudohomologues and (ii) a C25 head-to-tail linked (regular) isoprenoid hydrocarbon [possibly derived from dialkyl glycerol diether lipids (DGDs)] and its degradation products, C21–24 pseudohomologues and abundant pristane and phytane. Through combination of compound-specific stable carbon isotope analysis of isoprenoid hydrocarbons, which had average δ13C values of −35‰ to −30‰, and their molecular distribution, it was not possible to unambiguously define the archaeal source for the biphytanes in the microbialite. The δ13C values for pristane and phytane were similar to those for head-to-tail linked C21–25 isoprenoids; potential source organisms for these compounds were halophilic archaea. Except for methane seep microbialites, no other ancient or recent phototrophic microbialites have been reported to contain predominantly archaeal isoprenoid hydrocarbons. Our findings suggest the presence of a new type of microbialite.

Evaluating degradation of hexachlorcyclohexane (HCH) isomers within a contaminated aquifer using compound-specific stable carbon isotope analysis (CSIA)

The applicability of compound-specific stable carbon isotope analysis (CSIA) for assessing biodegradation of hexachlorocyclohexane (HCH) isomers was investigated in a contaminated aquifer at a former pesticide processing facility. A CSIA method was developed and tested for efficacy in determining carbon isotope ratios of HCH isomers in groundwater samples using gas chromatography – isotope ratio mass spectrometry (GC-IRMS). The carbon isotope ratios of HCHs measured for samples taken from the field site confirmed contaminant source zones at former processing facilities, a storage depot and a waste dump site. The 13C-enrichment in HCHs provided evidence for biodegradation of HCHs especially downstream of the contaminant source zones. CSIA from monitoring campaigns in 2008, 2009 and 2010 revealed temporal trends in HCH biodegradation. Thus, the impact and progress of natural attenuation processes could be evaluated within the investigated aquifer. Calculations based on the Rayleigh-equation approach yielded levels of HCH biodegradation ranging from 30 to 86 %. Moreover, time- and distance-dependent in situ first-order biodegradation rate constants were estimated with maximal values of 3 × 10−3 d−1 and 10 × 10−3 m−1 for α-HCH, 11 × 10−3 d−1 and 37 × 10−3 m−1 for β-HCH, and 6 × 10−3 d−1 and 19 × 10−3 m−1 for δ-HCH, respectively. This study highlights the applicability of CSIA for the assessment of HCH biodegradation within contaminated aquifers.